Objective To investigate the characteristics of human immunodeficiency virus(HIV-1)V3 loop amino acid mutations among AIDS patients in Shenzhen. Methods Fragments of HIV-1 env gene were amplified by RT-nested-polymerase chain reaction(PCR)from plasma of the AIDS patients in Shenzhen.The PCR products were then directly sequenced and the sequences were aligned,translated and analyzed. Results Sequence analysis showed that there were B and CRF01-AE HIV-1 subtypes in Shenzhen.The V3 loop terra peptide GPGQ accounted for about 48%,GPGR about 36%,other V3 loop terra peptide forms were 16%;the V3 loop central motif in the majority(76.9%)of the CRF01-AE strains was GPGQ,the majority(75%)of the B strains was GPGR.We also found some unusual terra peptide compositions:DQDR and DQGQ on the tip of V3 loop,some amino acid mutations at specific of V3 loop associated with SI phenotype were very common in AIDS patients(80%). Conclusions The V3 loop amino acid mutations were very common in AIDS patients,the V3 loop central motif in the majority was GPGQ and GPGR in Shenzhen,the mutations shows the change of HIV-1 phenotype and predicts that disease progress to AIDS.

The construction of a network resource data base for clinical skill teaching aiming at sharing excellent teaching resources was completed by integrating all kinds of teaching resource including characters,eourseware,pictures,cartoons,videos and examination questions,etc.In view of the generally weak situation of the current domestic education resource data base construction,the contents,guiding ideology,principle,object,orientation,function and the management mechanism of the network resource data base construction were devised for clinical skills teaching.Suggestions for specific ideas and construction problems were proposed to promote the construction and optimization of the network resource database for clinical skills teaching.

In order to construct generally efficient cell immortalization vector, pTP-hTERT, we modified the traditional piggyBac (PB) transposon using artificial synthesis, PCR and enzyme digestion. The modified vector contained the necessary transposon elements, a PB transposase expression cassette, a co-expression selectable element and a human telomerase reverse transcriptase (hTERT) expression cassette. The co-expression selectable element had two markers, enhanced green fluorescent protein (EGFP) gene and puromycin-resistance (Puro) gene, linked by porcine teschovirus-1 2A peptide (P2A). To validate the functionality of vector elements, we transfected pTP-hTERT into HEK293 cell, selected the positive cell clones and then conducted RT-PCR, Western blotting (WB) and Tail-PCR, methylene blue staining and statistic analysis on selected cells. The results of sequencing and cell culture show that the pTP-hTERT was constructed successfully and the positive cell could be selected by puromycin. The WB results, P2A cutting EGFP and Puro fusion protein with high efficiency, reflected the selectable element worked. The sequencing result of Tail-PCR confirmed the vector integrated into the genome through transposition. The results of methylene blue staining and statistic analysis indicated the clone of positive cells triggered by pTP-hTERT significantly increased (P < 0.01) compared with control group. The construction of pTP-hTERT provides an efficient tool for establishing immortalized cell lines and a demonstration for building other eukaryotic plasmids.
Cell Line ; DNA Transposable Elements ; genetics ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; HEK293 Cells ; Humans ; Plasmids ; Polymerase Chain Reaction ; Telomerase ; genetics ; Transfection

OBJECTIVE: To determine the feasibility whether the bovine jugular venous conduit (BJVC) can be fixed with polyepoxy compound (PC). METHODS: Twenty-four BJVCs were divided into 3 groups and fixed with polyepoxy compound (PC group, n = 8), glutaraldehyde (GA group, n = 8), and unfixed group (Control group, n = 8), respectively. The morphologic and mechanical properties of BJVCs in the 3 groups, including thickness, diameter, moisture content, denaturation temperature, tensile strength, elongation at break, and fixation index were measured. The rat subcutaneous model for the assessment of tissue calcification was used. The calcium content in bovine jugular vein patches and valves was determined by flame atomic absorption spectrophotometer. RESULTS: There was no difference in the wall thickness, diameter, and tissue water content between PC and the control group, but significant difference was found between GA and PC groups. The mechanical properties of PC group and GA group were not significantly different, but they were better than those of the control group. GA-fixed BJVC samples showed clear calcification, while PC fixed BJVC were calcified significantly less. CONCLUSION PC is an effective and suitable choice for the treatment of BJVC since it can effectively preserve the structure and the anti-reflow function of valves in bovine jugular vein and it has better anti-calcification properties.
Animals ; Biocompatible Materials ; Bioprosthesis ; Blood Vessel Prosthesis ; Cattle ; Cross-Linking Reagents ; pharmacology ; Epoxy Compounds ; pharmacology ; Jugular Veins ; Polymers

OBJECTIVETo study immunomodulating activity of Lonicera Japonica flavone by investigating immune enzymatic activity of serum and antoxidized activity of lymphoid organs in mice.

METHODSFifty KM mice were randomly divided into control group, model group, low dose group, middle dose group and high dose group(n = 10), respectively. And low dose group, middle dose group and high dose group were given Lonicera Japonica flavone with 100 mg/kg, 200 mg/kg and 400 mg/kg every day, respectively, while control group and model group were administered with NS. After continuously giving drug 7 weeks, other groups were injected with Dexamethasome (Dex: 25 mg /kg) for 3 days by subcutaneous injection, but the control group were treated with NS. And after giving Lonicera Japonica flavone 1 week simultaneously, organ indexes , the activity of acid phosphatase (ACP), alkaline phosphatase (AKP) and lysozyme (LSZ) in serum , and the content of monoamine oxidase (MAO), total antioxidant capacity (T-AOC), total superoxide dismutase (SOD) and malondialdehyde (MDA) in lymphoid organs in mice were tested, respectively.

RESULTSLonicera Japonica flavone could significantly improve the organ indexes, and significantly improve the activity of ACP, AKP and LSZ in serum, and significantly improve the contents of T-AOC and SOD, but reduce that of MAO and MDA in lymphoid organs in immunosuppressed mice.

CONCLUSIONIonicera Japonica flavone can significantly improve the activity of immune enzyme in serum and the antioxidized activity of lymphoid organs in mice. It suggests that Ionicera Japonica flavone has a good immunomodulatory effects.

Acid Phosphatase ; blood ; Alkaline Phosphatase ; blood ; Animals ; Antioxidants ; metabolism ; Flavones ; pharmacology ; Immunomodulation ; Lonicera ; chemistry ; Malondialdehyde ; metabolism ; Mice ; Monoamine Oxidase ; metabolism ; Muramidase ; blood ; Superoxide Dismutase ; metabolism

AIM: To investigate the effects of Yiqi Huoxue prescription and Bushen Shengsui prescription on the expression of Notch-1 and Jagged1 protein in ischemic penumbra after local cerebral ischemia/reperfusion in intraluminal thread occlusion of the middle cerebral artery (MCAO) rats. METHODS: The local cerebral ischemia/reperfusion model was established by intraluminal thread occlusion of the middle cerebral artery. The animals were randomly divided into pseudo surgery group (sham group), model (MCAO) group, Yiqi Huoxue prescription (MCAO+Yiqi Huoxue) and Bushen Shengsui prescription (MCAO+Bushen Shengsui) groups. Using the techniques of immuno-histochemical staining, the expression of Notch-1 and Jagged1 was observed at 1, 2 and 3 weeks in ischemic penumbra of the cortex of frontal and parietal lobe. RESULTS: A value and positive unit of Notch-1 and Jagged1 protein expression in model group were higher than those in sham in ischemic penumbra of the cortex of frontal or parietal lobe after local cerebral ischemia/reperfusion in MCAO rats (P<0.05 or P<0.01). A value and/or positive unit of the expression in Yiqi Huoxue prescription and Bushen Shengsui prescription groups were lower than those in model group (P<0.05 or P<0.01). CONCLUSION: Yiqi Huoxue prescription and Bushen Shengsui prescription affect the proliferation and differentiation of neural stem cells, depress Notch signal transduction after local cerebral ischemia/reperfusion in MCAO rats by inhibiting the expression of Notch-1and Jagged1 protein.

Objective To investigate the correlation between the apparent diffusion coefficient (ADC) and the expression of aquaporin-4 (AQP4) in brain tissue after ischemia/reperfusion in rats. Methods A model of right middle cerebral artery occlusion (MCAO) induced by suture method. Seventy Wistar rats were randomly divided into 7 groups: sham-operation (A), MCAO 30 min (B), MCAO 30 min and reperfusion 30 min (D), MCAO 30 minand reperfusion 60 min (E), MCAO 60 min (C); MCAO 60 min and reperfusion 30 min (F), and MCAO 60 min and reperfusion 60 min (G) groups (n=10 in each group). The rats in all groups underwent diffusion-weighted imaging (DWI). The relative apparent diffusion coefficient (rADC) was calculated. Triphenyltetrazolium chloride (TTC) staining was used to detect the ratio of ischemic area. Immunohistochemistry, in site hybridization, and reverse transcription-polymerase chain reaction (RT-PCR) were used to detect the expressions of AQP4. Results No abnormal intensity signals were observed on DWI in sham-operation group. The ranges of the high signal intensity lesions on diffusion-weighted imaging (DWI) were from small to large in groups B, C, D, E, F, and G. Then it reduced gradually, so did the ischemic area. The AQP4 expression was up-regulated significantly in groups B and E. The AQP4 expression was decreased significantly after the reperfusion in all groups. 1here was significant difference between ischemic groups and reperfusion groups (all P<0. 05). The expressions of rADC and AQP4 showed negative correlation (r=0. 72, P<0.01). Conclusions The AQP4 expression in brain tissue is closely associated with the changes of ADC after cerebral ischemia/reperfusion. DWI may indirectly reflect the levels of AQP4 expression.

Objective To discuss the diagnosis and treatment of cerebral sparganosis. Methods To summary four cases of cerebral sparganosis, focusing on the clinical course and imaging findings, with the goal of better diagnostic skills. Results All 4 cases had some kind of misdiagnosis and improvement after surgery or parasiticidal pharmacotherapy. Cerebral MRI and CT scans revealed relatively extensive white matter degeneration and focal enhancements. Subsequent scans showed changes in shape and location of the enhanced foci, indicating the migration of sparganum. Pathologic findings of 3 patients who had undergone surgery showed granuloma and sparganum. Conclusions Cerebral sparganosis has relatively special manifestions on imaging, which are of diagnostic value. The spaganum should be as completely removed as possible during surgery.

Objective To explore the clinical significance of N-terminal pro-brain natriuretic peptide(NT-proBNP)in the evaluation of cardiac function in patients with severe burns. Methods A total of 78 patients with severe burns in intensive care unit(ICU)in our hospital were selected. Patients were divided into a heart failure group(n = 31)and a non-heart failure group(n = 47)according to the heart failure complication condition. The changes of NT-proBNP level in different time ,heart function index ,relevant index and mortality were compared between the two groups. Results The levels of NT-proBNP ,troponin(CTnT),creatine kinase(CK),creatine kinase isoenzyme (CK-MB),stroke volume index (SVI),cardiac index (CI) and central venous pressure (CVP),and the score of acute physiologic and chronic health conditions(APACHE II)in the heart failure group were shown to be significantly increased than those of the non-heart failure group (P < 0.05). The mean arterial pressure(MAP)in the heart failure group was significantly lower than the non-heart failure group(P < 0.05). Mortality rate in the heart failure group was shown to be significantly higher than the non-heart failure group (P < 0.05). Conclusion Heart failure is a common complication in patients with severe burns. The mortality rate in the population with heart failure is relatively high ,for which NT-proBNP can effectively assess the conditions and prognosis.

This study aims to analyse the value of CODEHOP RT-PCR in the detection of Flavivirus. According to the amino acid sequences of polyproteins of different flaviviruses published in GenBank, a pair of primers was designed using the CODEHOP method. One-step RT-PCR was used to detect Japanese encephalitis virus strain JEV1201, Dengue virus strain JKD001, and yellow fever virus vaccine YV6161. BLAST analysis and phylogenetic analysis were performed after the RT-PCR products of nucleocapsid genes were sequenced. The results showed that this method could amplify Flavivirus specifically, and the size and sequence of the target fragment accorded with the anticipated result. JEV1201 had the highest homology to Japanese encephalitis virus strain YL2009-4/YC2009-3, belonging to the branch of the phylogenetic tree of Japanese encephalitis virus strains. JKD001 had the highest homology to Dengue virus strain DENV-2/ID/1022DN/1975, belonging to the branch of the phylogenetic tree of Dengue virus strains. YV6161 had the highest homology to Yellow fever virus strain 17D, belonging to the branch of the phylogenetic tree of Yellow fever virus strains. In conclusion, the method of CODEHOP RT-PCR can be effectively used to detect, identify, and phylogenetically analyse Flavivirus.
DNA Primers ; genetics ; Flavivirus ; classification ; genetics ; isolation & purification ; Flavivirus Infections ; virology ; Humans ; Molecular Sequence Data ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Viral Nonstructural Proteins ; genetics